Plasma membrane fluidity measurements on whole living cells by fluorescence anisotropy of trimethylammoniumdiphenylhexatriene

Abstract

Trimethylammoniumdiphenylhexatriene (TMA-DPH) is a hydrophobic fluorescent probe with a high quantum yield, which was shown earlier to have specific localization properties in the plasma membranes of whole living cells. This probe was used in aqueous suspensions of L929 mouse fibroblasts, rat mast cells and ReH6 leukemic lymphocytes for determining plasma membrane fluidity from fluorescence stationary anisotropy measurements. TMA-DPH was only partially incorporated into the membranes, most of it remained as a stable form in the buffer solution; the distribution was governed by an equilibrium. The measurements were influenced by unavoidable parasitic scattered light and an appropriate correction is described. A set of precautions for the proper use of the probe is proposed. The results indicated that the fluidity was considerably lower in whole cells than in isolated membranes from the same system.